EP1412346A1

EP1412346A1 - Method for producing novel spinosyn derivatives

Info

Publication number: EP1412346A1
Application number: EP02754847A
Authority: EP
Inventors: Peter Jeschke; Günther EBERZ; Rita FRÖDE; Volker MÖHRLE; Robert Velten
Original assignee: Bayer CropScience AG
Current assignee: Bayer CropScience AG
Priority date: 2001-07-20
Filing date: 2002-07-08
Publication date: 2004-04-28
Anticipated expiration: 2022-07-08
Also published as: DE10135550A1; US20060173172A1; CN1293069C; WO2003010155A1; US20040242858A1; JP4451132B2; JP2005503371A; US7034130B2; ATE290532T1; CN1556802A; DE50202431D1; EP1412346B1

Abstract

The invention relates to a method for producing novel spinosyn derivatives, which are substituted in the C-21 position by a 1-hydroxy-ethyl radical, to novel spinosyn derivatives of this type per se and to their use for producing novel spinosyns.

Description

Process for the preparation of new spinosyn derivatives

The present invention relates to processes for the preparation of new spinosyn derivatives which are substituted in the C-21 position with a 1-hydroxyethyl residue, and to such new spinosyn derivatives per se and their use for producing new spinosyns.

The spinosyns are known compounds. Spinosyns are fermentation products made from cultures of the Actinomycete Saccharopolyspora spinosa. Natural spinosyns consist of a tetracyclic polyketide backbone (aglycon) with a 12-membered macrolide ring and a 5,6,5-cis-anti-trans-tricyclic ring, as well as a D-forosamine and a 2,3,4-tri-O- methyl L-rhamnose sugar content (Kirst et al., 1991, Tetrahedron Letters, 32: 4839). More than 20 different natural spinosyns, the so-called A83543 complex, have been described so far (cf. WO 97/00265, WO 94/20518 and WO 93/09126). For example, EP-A 375316 describes the spinosyns A, B, C, D, E, F, G, H and J. Spinosyns L, M, N, Q, R, S and T are known from WO 93/09126. WO 94/20518 mentions the spinosyns K, O, P, U, V, W and Y and their derivatives. These compounds vary in the substitution of one or a few methyl groups on the tetracyclic backbone, on the D-forosamine or on the 2,3,4-tri-O-methyl-L-rhamnose sugar portion. A 17-pseudoaglycone lacking the D-forosamine sugar content has also been isolated from culture broths from S. spinosa.

The main components of the A83543 complex formed by S. spinosa are the variants Spinosyn A and Spinosyn D, which are the essential components of the product Spinosad (see Pesticide Manual, British Crop Protection Council, 1 ^lth Ed., 1997, page 1272 and Dow Elanco Trade Magazine Down to Earth, Vol. 52, NO. 1, 1997 and the literature cited therein). If the amino sugar is not present at the C-17 position, the compounds are referred to as spinosyn A, D, etc.-17 pseudoaglycone; if the neutral sugar is not present in the C-9 position, the compounds are referred to as spinosyn A, D, etc.-9 pseudoaglycone. Spinosyns without the two sugar residues in positions C-9 and C-17 are called spinosyn aglycone.

Spinosyns are suitable for controlling arachnids, nematodes, ectoparasites (cf. WO 01/11962, WO 01/11963, WO 01/11964) and insects, in particular of lepidopterans and dipteran species. In addition, the technical application of Spinosyne is environmentally friendly and this class of substances also has an attractive toxicological profile.

However, it can be expected that animal pests, in particular ectoparasites or plant pests, which are currently being combated with spinosyns, can develop resistance to these active substances which are on the market.

It is therefore important to produce new biologically active derivatives of spinosyns that can replace the spinosyns currently used to control pests.

Certain natural aglycone derivatives of spinosyn that contain a hydroxyl group

C-8 position of the macrolide backbone have recently been using Saccharopolyspora sp. (LW107129) and have become known as insecticidally active compounds (cf. WO 01/19840). While numerous modifications to spinosyns were carried out (cf. WO 97/00265), derivatization ranges on the methyl and ethyl groups in the C-21 position of the macrolide backbone received little attention. Although the functionalization of the alkyl radical in the C-21 position would be advantageous, inter alia, for derivatization reactions, only a few spinosyn derivatives are known whose substituents in C-21 have a hydroxyl group. For example, spinosyn derivatives which have been substituted with a 3-hydroxy-1-butenyl radical in C-21 have recently been described, optionally additionally can carry a hydroxyl group in the above-mentioned C-8 position and have an insecticidal activity (cf. WO 01/19840).

While chemically synthetic processes for the preparation of spinosyn derivatives have been described (cf. Martynow, J.G. and Kirst, H.A., 1994, J. Org. Chem., 59:

1548), nothing is known about the chemical synthesis of the above-mentioned spinosyn derivatives with a 1-hydroxyethyl residue in the C-21 position.

It is the object of the present invention to provide suitable processes which are suitable for the selective and / or stereospecific production of new spinosyn-

Derivatives with a 1-hydroxyethyl residue in the C-21 position can be used.

The object was achieved by the provision of processes for the preparation of compounds of the general formula (I),

in which

AB is one of the following groups: -HC = CH-, -HC = C (CH ₃ ) -, -H ₂ C-CH ₂ - or -H ₂ C-CH (CH ₃ ) -,

D for the group

or stands R ^{1 represents} hydrogen or an amino sugar, and

R represents hydrogen or a sugar,

where compounds of the general formula (II),

in which

AB, D and R ^{1 have} the meanings given above,

with a microorganism in an aqueous nutrient medium under aerobic conditions or with an enzyme extract produced therefrom or with one or more enzymes isolated therefrom.

The starting compounds are thus converted selectively and / or stereospecifically by biotransformation using microorganisms or their enzymes into spinosyn derivatives which are substituted in the C-21 position with a 1-hydroxyethyl residue.

The term "spinosyn derivatives" as used herein also includes spinosyn-aglycone compounds, ie compounds which have the macrolide backbone of the spinosyns but no sugar residues. Those compounds of the general formula (IT) are preferably used as starting compounds,

in which case (1) that

AB stands for one of the following groups: -HC = CH-, -HC = C (CH ₃ ) -, -H ₂ C-CH ₂ - or -H ₂ C-CH (CH ₃ ) -, and

D for the group

stands,

R ^{1 represents} an amino sugar of the formula Ia,

1a and

R represents a sugar of the formula 2a,

me

2a

or in which case (2) that AB stands for the group -HC = CH- or -H ₂ C-CH ₂ -, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the above formula la, and

R ^{2 represents} hydrogen or a sugar of the formula 2b, 2c, 2d, 2e or 2f,

me

2 B

me

2e

me

2f

or in which case (3) that

A-B stands for one of the following grapples: -HC = CH-, -HC = C (CH) - or

-H ₂ C-CH ₂ -, and

D has the meaning given above,

R ^{1 represents} hydrogen or an amino sugar of the formula Ib,

1 bunch

R ^{2 represents} hydrogen or a sugar of the above formula 2a,

or in which case (4) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the above formula la, and R ι2 represents a sugar of the formula 2g, 2h, 2i, 2j or 2k,

me

2g

me

2h

me

2i

me

2k

or in which case (5) that

AB stands for the group -HC = CH- or -H ₂ C-CH ₂ -, and D has the meaning given above,

R ^{1 represents} an amino sugar of the above formula la, and

R represents a sugar of the formula 21 or 2m,

me

21

me

2m

or in which case (6) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and

D has the meaning given above,

R ^{1 represents} hydrogen or an aminosugar of the abovementioned formula lb or an aminosugar of the formula lc,

1c and

R 'represents a sugar of the above-mentioned formula 2b, 2c, 2g or 2h, or a sugar of the formula 2n,

me

2n

or in which case (7) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 stands} for an amino sugar of the above formula 1 a, and

R represents a sugar of the formula 20,

me

2o

or in which case (8) that

AB stands for the group -HC = CH-, and D has the meaning given above,

R ^{1 represents} an aminosugar of the abovementioned formula Ib, and

R represents a sugar of the above-mentioned formula 2d, 2i or 2j, or a sugar of the formula 2p,

2p

or in which case (9) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} hydrogen or an amino sugar of the above formula Ic, and

R ^{2 represents} a sugar of the above formula 2i or 2p,

or in which case (10) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the formula 1 d, 1 e or 1 f,

1d

1e

1f and

R ^{2 represents} a sugar of the abovementioned formula 2a,

or in which case (11) that

A-B stands for Grappe -HC = CH-, and

D stands for grappa, R ^{1 represents} hydrogen or an amino sugar of the above formula la.

Compounds of the general formula (II) which are particularly preferably used as starting compounds are

in which case (12) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and

D for the Grappe

R ^{1 represents} an amino sugar of the formula Ia, and

R ^{2 represents} a sugar of the formula 2a, 2g or 2h,

or in which case (13) that

A-B stands for Grappe -HC = CH-, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the formula 1a, and

R ^{2 represents} hydrogen or a sugar of the formula 2d, 2e, 21, 2m or 2o,

or in which case (14) that AB stands for the group -HC = CH- or -HC = C (CH ₃ , and

D has the meaning given above,

R ^{1 represents} hydrogen or an amino sugar of the formula Ib, and

R ^{2 represents} hydrogen or a sugar of the formula 2a,

or in which AB, D and R ^{1 are defined} as in case (11).

Very particular preference is given to using compounds of the general formula (IT) as starting compounds

in which case (15) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and

D for the group

stands,

R ^{1 represents} an amino sugar of the formula 1 a, and

R ^{2 represents} a sugar of the formula 2a,

or in which case (16) that

A-B stands for the group -HC = CH-, and

D has the meaning given above, R ^{1 represents} an amino sugar of the formula Ia, and

R ^{2 represents} hydrogen or a sugar of the formula 2d, 21 or 2m,

or in which AB, D and R ^{1 are defined} as in case (11).

Most preferably, compounds of the general formula (II) are used as starting compounds

in which case (17) that

AB stands for Grappe -HC = CH- or -HC = C (CH ₃ ) -, and

D for the group

stands,

R ^{1 represents} an amino sugar of the formula 1 a, and

R ^{2 represents} a sugar of the formula 2a,

or in which case (18) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} hydrogen, and

R ^{2 represents} hydrogen. The present invention also relates to the compounds of the general formula (I), in which AB, D and R ^{1 have} the meanings mentioned above.

The abbreviation "Me" used herein means methyl; the abbreviation "Et" stands for ethyl.

Both the optically active, stereoisomeric forms and also the diastereomeric forms of the compounds of the general formula (I) can be formed by the process according to the invention.

The compounds of formula (I) according to the invention can exist in stereoisomeric forms which either behave like image and mirror image (enantiomers) or which do not behave like image and mirror image (diastereomers). The present invention relates to both the enantiomers and the diastereomers and their respective mixtures. The racemic shapes can be just like that

Separate diastereomers in a known manner into the stereoisomerically uniform constituents. If appropriate, the isomers can be converted into one another by methods known per se.

The compounds of the invention, in which R ¹ for an aminosugar

Formulas l - le can form salts. Salts are formed according to standard salt manufacturing processes. For example, the compounds according to the invention are neutralized with appropriate acids in order to form acid addition salts. Representatively usable acid addition salts are salts which are formed, for example, by a reaction with other inorganic acids, such as for example sulfuric acid, hydrochloric acid, hydrobromic acid, phosphoric acid or organic carboxylic acids such as acetic acid, trifluoroacetic acid, citric acid, succinic acid, lactic acid, formic acid, maleic acid, camphic acid, phthalic acid, phthalic acid, phthalic acid Glutaric acid, stearic acid, salicylic acid, sorbic acid, cinnamic acid, picric acid, benzoic acid or organic sulfonic acids such as methanesulfonic acid and Form para-toluenesulfonic acid or with basic amino acids such as aspartic acid, glutamic acid, arginine or the like.

The compounds of the general formula (II) which can be used as starting compounds for the process according to the invention are known (cf. Creemer L.C. et al.,

1998, J. Antibiotics 51 (8): 795-800; Sparks T.C. et al., 1998, J. Econ. Entomol. 91 (6): 1277-1283; Sparks, T.C. et al., 2000, Pestic. Biochem. Physiol. 67 (3): 187-197; Paquette L.A. et al., 1998, J. Am. Chem. Soc. 120 (11): 2553-2563; Evans D.A. et al., 1993, J. Am. Chem. Soc. 115 (11): 4497-4513; Crouse G.D. et al., 2001, Pest. Manag. Be. 57 (2): 177-185; Creemer L.C. et al., 2000, J. Antibiotics 53 (2):

171-178; Sparks T.C. et al., 2000, Proc.-Beltwide Cotton Conf. Vol. 2: 1225-1229) or can be prepared by the process described in WO 01/16303. The starting compounds which can be used for the process according to the invention can be obtained analogously, starting from the corresponding natural spinosyns.

If, for example, the spinosyn A aglycone of the formula (Ha) is used, the process according to the invention can be reproduced by reaction scheme 1:

Reaction scheme 1

[Biotransformation]

(La)

Selective and / or stereospecific hydroxylations of natural products and synthetic compounds by bioconversion using microorganisms or their enzymes are described in the literature. In particular, cells and / or enzymes from Gram-positive bacteria, such as Streptomycetes, or Gram-negative bacteria, such as Pseudomonas, or fungi, such as Fusarium, have been used. Examples of microorganisms that contain corresponding enzyme classes, such as P-450 monooxygenases, are listed in the following table:

According to the invention and in accordance with reaction scheme 1 mentioned above, compounds of the general formula (11), in which AB, D and R ^{1 have} the meanings given above, can be brought into contact with a suitable microorganism in an aqueous nutrient medium under aerobic conditions and then the desired compounds isolate the general formula (I).

Instead of the microorganisms, it is also possible to use enzyme extracts and purified enzymes, optionally after adding or regenerating the required cofactors, which can be obtained from these microorganisms by conventional methods.

In a special way, genes which determine the biosynthesis of such enzymes can also be cloned and expressed in foreign hosts, such as, for example, Escherichia coli. Such recombinant bacteria can be used directly for biotransformation. In addition, an enzyme extract of such a recombinant cell or a purified protein, optionally after adding or regenerating the required cofactors, can be used for the biotransformation. A microorganism from the Grappe of the Actinomycetes, in particular from the genus Streptomyces, is preferably used for the process according to the invention.

A strain from the species Streptomyces djakartensis, Streptomyces griseofuscus, Streptomyces caelestis, Streptomyces antibioticus, Streptomyces griseus or Streptomyces aureofaciens is particularly preferably used for the method according to the invention.

A strain with the characteristic features of the following strains is very particularly preferably used for the method according to the invention:

The strains mentioned in the table have once again been deposited with the German Collection of Microorganisms and Cell Cultures GmbH (DSMZ), Mascheroder Weg lb, D-38124 Braunschweig, Germany, in accordance with the provisions of the Budapest Treaty:

The strains are described in more detail in Example 16. Not only the deposited strains as such, but also mutants thereof can be used as long as these mutants have the characteristic features of the deposited strains. This means that the mutants must still have the ability to be able to carry out the bioconversion according to the invention.

The aqueous nutrient medium preferably contains an assimilable carbon source and an assimilable nitrogen source.

The compounds of formula (I) are produced, for example, when a strain from the species Streptomyces djakartensis, S. griseofuscus, S. caelestis, S. antibioticus, S. griseus or S. aureofaciens in an aqueous nutrient medium under aerobic conditions in the presence of Compounds of formula (11) are fermented. Typically, the microorganisms are fermented in a nutrient medium which contains a carbon source and optionally a proteinaceous material. Preferred carbon sources include glucose, brown sugar, sucrose, glycerin, starch, corn starch, lactose, dextrin, molasses etc. Preferred nitrogen sources include cottonseed flour, yeast, autolysed

Baker's yeast, solid milk components, soybean meal, corn meal, pancreatic or papainically digested casein breakdown products, solid distillation components, animal peptone broths, meat and bone pieces, etc. Preferably combinations of these carbon and nitrogen sources are used. Trace elements such as zinc, magnesium, manganese, cobalt, iron etc. do not have to be added to the cultivation medium as long as tap water and non-purified components are used as medium components.

The preparation of the compounds of the general formula (I) can be carried out in any

Temperature are induced, which ensures sufficient growth of the microorganisms. The temperature is preferably between 21 ° C. and 32 ° C., particularly preferably approximately 28 ° C. In general, optimal production of the compounds of formula (I) is obtained within 2 to 4 days after adding the compounds of formula (II) to the culture. The compounds according to the invention can be produced both in shake bottles and in stirred fermenters.

Preferred growing conditions and media for growing in the shake flask are described in Examples 2 to 9.

The compounds according to the invention as a biotransformation product can be isolated from the cultivation medium by customary methods.

Various methods can be used to make the invention

Isolate and purify compounds from the fermentation broth, e.g. preparative gel chromatography, preparative chromatography on reversed phase or preparative absorption chromatography. The detection can e.g. by UV absorption or mass spectrometry.

The compounds according to the invention can be used for the production of biologically active, in particular insecticidal and acaricidal, spinosyns. The biological effectiveness of certain natural aglycone derivatives of spinosyn which have a hydroxyl group in the C-8 position of the macrolide structure has recently become known (cf. WO 01/19840). As further examples, the spinosyn derivatives with a 3-hydroxy-1- To name butenyl radical in the C-21 position. If appropriate, these spinosyn derivatives can additionally carry a hydroxyl group in one of the above-mentioned C-8 positions and also have an insecticidal activity (cf. WO 01/19840).

In the preparation of further spinosyn derivatives from the compounds according to the invention of the general formula (I) in which R ^{1 is} hydrogen and D is Grappe C = O or COR, in which R is hydrogen, it is advantageous to use suitable protective groups (SG) to use. For example, protective groups (SG) for hydroxyl groups, substituted methyl ethers and ethers, are substituted

Ethyl ethers, substituted benzyl ethers, silyl ethers, esters, carbonates or sulfonates are known (see Greene TW, Wuts PGW in Protective Groups in Organic Synthesis; John Wiley & Sohns, Inc. 1999, Protection for the hydroxyl group, including 1,2- and 1 , 3-diol).

Examples of protective groups (SG) of the substituted methyl ether type which may be mentioned are: methoxymethyl ether (MOM), methylthiomethyl ether (MTM), (phenyldimethylsilyl) methoxymethyl ether (SMOM-OR), benzyloxymethyl ether (BOM-OR), para-methoxybenzyloxymethyl ether (PMBM- OR), para-nitrobenzyloxy methyl ether, ortho-nitrobenzyloxymethyl ether (NBOM-OR), (4-methoxyphenoxy) methyl ether (p-AOM-OR), guaiacol methyl ether (GUM-OR), tert-butoxymethyl ether, 4-pentenyl -oxymethyl ether (POM-OR), silyloxymethyl ether, 2-methoxy ethoxymethyl ether (MEM-OR), 2,2,2-trichloroethoxymethyl ether, bis (2-chloroethoxy) methyl ether, 2- (trimethylsilyl) ethoxymethyl ether (SEM -OR), methoxy methyl ether (MM-OR).

Examples of protective groups (SG) of the substituted ethyl ether type which may be mentioned are: 1-ethoxyethyl ether (EE-OR), l- (2-chloroethoxy) ethyl ether (Cee-OR), l- [2- (trimethylsilyl) ethoxy] ethyl ether ( SEE-OR), 1-methyl-1-methoxyethyl ether (MTP-OR), 1-methyl-1-benzyloxyethyl ether (MBE-OR), 1-methyl-1-benzyloxy-2-fluoro-ethyl ether, 1-methyl-1 -phenoxy-ethyl ether, 2,2,2-trichloroethyl ether, 1,1-dianisyl- 2,2,2-trichloroethyl ether (DATE-OR), 1,1,1, 3,3,3-hexafluoro-2-ρhenyliso-ρroρyl ether (HIP-OR), 2-trimethylsilylethyl ether, 2- (benzylthio) ethyl ether , 2- (phenyl-selenyl) ethyl ether. Further protective groups (SG) of the ether type which may be mentioned are, for example: tetrahydropyranyl ether (THP-OR), 3-bromo-tetrahydropyranyl ether (3-BrTHP-OR), tetrahydrothiopyranyl ether, 1-methoxycyclohexyl ether, 2- and 4-

Picolyl ether, 3-methyl-2-picolyl-N-oxido ether, 2-quinolinyl methyl ether (Qm-OR), 1-pyrenyl methyl ether, dipenyl methyl ether (DPM-OR), para, para'-dinitrobenzohydyl ether (RO-DNB ), 5-dibenzosuberyl ether, triphenyl methyl ether (Tr-OR), α-naphthyl diphenyl methyl ether, para-methoxy-phenyldiphenyl methyl ether (MMTr-OR), di (para-methoxy-phenyl) phenyl methyl ether (DMTr-OR), tri (para-methoxy-phenyl ) methyl ether (TMTr-OR), 4- (4'-bromo-ρhenacyloxy) phenyldiphenylmethyl ether, 4,4 ', 4 "tris (4,5-dichlorophthalimido-ρhenyl) methyl ether (CPTr-OR), 4,4 ', 4 "- tris (levulinoyloxy-phenyl) methyl ether (TLTr-OR), 4,4', 4" - tris (benzoyloxyphenyl) - methyl ether (TBTr-OR), 4,4'-dimethoxy-3 '' - [ N- (imidazolylmethyl)] - trityl ether (IDTr-OR), 4,4'-dimethoxy-3 "- [N- (imidazolyl-ethyl) carbamoyl] trityl ether (IETr-

OR), l, l-bis (4-methoxyphenyl) - pyrenyl methyl ether (Bmpm-OR), 9-anthryl ether, 9- (9-phenyι) xanthenyl ether (pixyl-OR), 9- (9-phenyl -10-oxo) anthryl (tritylon ether). 4-methoxy-tetrahydropyranyl ether (MTHP-OR), 4-methoxy-tetrahydrothiopyranyl ether, 4-methoxy-tetrahydrothiopyranyl ether-S, S-dioxide, l - [(2-chloro-4-methyl) phenyl] -4- methoxypiperidin-4-yl ether (CTMP-OR), 1- (2-fluorophenyl) -4- methoxy-piperidin-4-yl ether (Fpmp-OR), 1,4-dioxan-2-yl ether, Tetrahydrofuranyl ether, TelahyάrotMofuranylether, 2,3,3a, 4,5,6, 7,7a-Octahydro-7,8,8-trimethyl-4,7-methanbenzofuran-2-yl ether (RO-MBF ), tert-butyl ether, allyl ether, propargyl ether, para-chlorophenyl ether, para-methoxyphenyl ether, para-nitrophenyl ether, 2,4-dinitro-phenyl ether (RO-DNP), 2,3,5,6-tetrafluoro-4- (trifluoromethyl) phenyl ether, benzyl ether (Bn-OR). Examples of protective groups of the substituted benzyl ether type are: para-methoxybenzyl ether (MPM-OR), 3,4-dimethoxy-benzyl ether (DMPM-OR), ortho-nitrobenzyl ether, para-nitrobenzyl ether, para-halobenzyl ether, 2,6-dichlorobenzyl ether, para-cyanobenzyl ether, para-phenyl benzyl ether, 2,6-difluorobenzyl ether, para-aminoacylbenzyl ether (PAB-OR), para-azidobenzyl ether (Azb-OR), 4-azido-3-chlorobenzyl ether, 2-trifluoromethyl- benzyl ether, para (methylsulfinyl) benzyl ether (Msib-OR). Examples of protective groups (SG) of the silyl ether type which may be mentioned are: trimethylsilyl ether (TMS-OR), triethylsilyl ether (TES-OR), triisopropylsilyl ether (TIPS-OR), dimethylisopropylsilyl ether (IPDMS-OR), diethylisopropylsilyl ether ( DEIPS-OR), dimethyl hexylsilyl ether (TDS-OR), tert-butyldimethylsilyl ether (TBDMS-OR), tert-butyl diphenylsilyl ether (TBDPS-OR), tribenzylsüyl ether, tri-para-xylylsilyl ether, triphenylsilyl ether (TPS- OR), diphenylmethylsilyl ether (DPMS-OR), di-tert-butylmethylsilyl ether (DTBMS-OR), tris (trimethylsilyl) silyl ether (sisyl ether), (2-hydroxystyryl) dimethylsilyl ether (HSDMS-OR), (2-hydroxystyryl) diisopropyl silyl ether (HSDIS-OR), tert-butyl methoxyphenylsilyl ether (TBMPS-OR), ter -

Butoxydiphenylsilyl ether (DPTBOS-OR). Examples of protective groups (SG) of the ester type include: formate ester, benzoyl formate ester, acetate ester (RO-Ac), chloroacetate ester, dichloroacetate ester, trichloroacetate ester, trifluoroacetate ester (RO-TFA), methoxy-acetate ester, triphenylmethoxyacetate ester, phenoxyacetate ester, phenoxyacetate ester, phenoxyacetate ester acetate esters, phenylacetate esters, diphenylacetate esters (DPA-OR),

Nicotinate ester, 3-phenylpropionate ester, 4-pentenoate ester, 4-oxopentanoate ester (levulinate) (Lev-OR), 4,4- (ethylenedithio) pentanoate ester (RO-LevS), 5- [3-bis (4-methoxyphenyl) hydroxy- methylphenoxy] levulinate ester, pivaloate ester (Pv-OR), 1-adamantanoate ester, crotonate ester, 4-methoxy-crotonate ester, benzoate ester (Bz-OR), para-phenyl-benzoate ester, 2,4,6-trimethylbenzoate ester (mesitoate), 4- (Methylthio-methoxy) butyrate ester (MTMB-OR), 2- (methylthiomethoxymethyl) benzoate ester (MTMT-OR). Examples of protective groups (SG) of the ester type include: methyl carbonate, methoxymethyl carbonate, 9-fluorenylmethyl carbonate (Fmoc-OR), ethyl carbonate, 2,2,2-trichloroethyl carbonate (Troc-OR), 1,1-dimefhyl-2, 2,2-trichloroethyl carbonate (TCBOC-OR), 2- (trimethylsilyl) ethyl carbonate

(TMSEC-OR), 2- (phenylsulfonyl) ethyl carbonate (Psec-OR), 2- (triphenylρhos-phonio) ethyl carbonate (Peoc-OR), tert-butyl carbonate (Boc-OR), isobutyl carbonate, vinyl carbonate, allyl carbonate ( Alloc-OR), p-nitrophenyl carbonate, benzyl carbonate (Z-OR), para-methoxybenzyl carbonate, 3,4-dimethoxybenzyl carbonate, ortho-nitrobenzyl carbonate, para-nitrobenzyl carbonate, 2-dansylethyl carbonate (Dnseoc-

OR), 2- (4-nitrophenyl) ethyl carbonate (Npeoc-OR), 2- (2,4-dinitrophenyl) ethyl carbonate (Dnpeoc-OR). Examples of protective groups (SG) of the sulfonate type which may be mentioned are: allylsulfonate (Als-OR), methanesulfonate (Ms-OR), benzyl sulfonates, tosylates (Ts-OR), 2 - [(4-nitrophenyl) ethyl] sulfonates (NPES-OR).

When producing further spinosyn derivatives from the above-mentioned compounds of the general formula (I), it may well be advantageous that the 1-hydroxyethyl radical in the C-21 position and optionally the hydroxyl group in the C-9 position (in the case of R ² = H) with suitable, for example with one of the above-mentioned protective groups (SG). The use of two different protective groups (SG ¹ or SG ² ) is recommended, whereby these should be compatible, ie they can be split off selectively and independently of one another. A derivatization, for example a glycosidation ring by means of chemical synthesis or by microbial bioconversion (cf. also US 5539089; introduction of the radicals R or R ') to the spinosyn derivatives (Ia-2 or Ib-2), on the hydroxyl group in the C-17 position take place (see scheme

2 and 3).

Scheme 2 - protecting group strategy (D: C-OH)

(la-3) (la-2)

(i) Introduction - protecting group (SG ¹ , SG ² ) (ii) derivatization - e.g. B. Glycosidation (R or R ')

(iii) Deblocking - protection group (SG) / or selective deblocking - protection group (e.g. SG ¹ ) Scheme 3 - protecting group strategy (D: C = O)

(Ib) (lb-1)

(i) Introduction - Protection Group (SG)

(ii) derivatization - e.g. B. Glycosidation (R)

(iii) Deblocking - Protection Group (SG)

After successful derivatization in positions C-9 or C-17 (see Ia-4) or C-17 (see Ib-2), the remaining protective group (SG) can be split off and that

Spinosyn derivative of the general formula (I) can be obtained.

Examples

Example 1 Strains Used

Table: Strains capable of biotransforming compound (fla) into the corresponding derivative with a 1-hydroxyethyl residue in the C-21 position (compound (la)):

ATCC = American Type Culture Collection, Manassas, VA, U.S.A.

DSM = German Collection for Microorganisms and Cell Cultures, Braunschweig,

Germany.

NRRL = Agricultural Research Service Culture Collection, Peoria, IL, U.S.A.

Example 2

Biotransformation with S. djakartensis NRRL B-12103

Exemplary protocol of the biotransformation for the production of the compound (la) from the compound (fla).

To prepare the production cultures, 20 ml of R5A medium (R5 A medium per liter: 103 g of sucrose; 0.25 g of K ₂ SO ₄ ; 10.12 g of MgCl ₂ ; 10 g of glucose; 5 g yeast extract; 0.1 g casamino acids, 21 g MOPS buffer pH 6.8 (KOH), 2 ml trace element solution; Trace element solution: (per liter) 40 mg ZnCl ₂ , 200 mg FeCl x 6 H ₂ O, 10 mg CuCl ₂ x 2H ₂ O, 10 mg MnCl ₂ x 4 H ₂ O, 10 mg Na ₂ BO ₇ x 10 H ₂ O , 10mg (NH ₄ ) ₆ Mo ₇ O ₂₄ x 4 H ₂ O; (Fernandez et al., 1998, J. Bacteriol. 180: 4929; Hopwood et al., 1985, Genetic manipulation of Streptomyces. A laboratory manual.

The John Innes Foundation, Norwich, England) in 100 ml Erlenmeyer flasks with 50 μl spore suspension of S. djakartensis NRRL B-12103. Before inoculation, the medium was sterilized at 121 ° C and 1.1 bar pressure for 20 minutes. The cultures were incubated at 28 ° C and 200 rpm. After 24 hours and after 72 hours of incubation, 1 mg of the compound (Ha) (100 μl each

Stock solution of 10 mg / ml in methanol) added. The biotransformation was stopped after 120 hours. The cultures were centrifuged off (10 minutes, 4000 rpm) and the supernatant was mixed with the same volume of methanol.

Example 3

Biotransformation with Streptomyces griseofuscus

Exemplary protocol of a biotransformation for the production of the compound (la) from the compound (Tla).

The method of Example 2 was used to prepare the production cultures using 50 μl spore suspension of the strain Streptomyces griseofuscus instead of the strain S. djakartensis NRRL B-12103.

Example 4

Biotransformation with Streptomyces caelestis

Exemplary protocol of a biotransformation for the production of the compound (la) from the compound (Ha).

The method of Example 2 was used to prepare the production cultures using 50 μl spore suspension of the strain Streptomyces caelestis instead of the strain S. djakartensis NRRL B-12103.

Example 5

Biotransformations with Streptomyces antibioticus

Exemplary protocol of a biotransformation for the production of the compound (Ia) from the compound (ITA).

The method of Example 2 was used to prepare the production cultures using 50 μl spore suspension of the Streptomyces antibioticus strain instead of the S. djakartensis NRRL B-12103 strain.

Example 6

Biotransformation with Streptomyces griseus

Exemplary protocol of a biotransformation for the production of the compound (la) from the compound (ITa).

The method of Example 2 was used to prepare the production cultures using 50 μl spore suspension of the strain Streptomyces griseus instead of the strain S. djakartensis NRRL B-12103. Example 7

Biotransformation with Streptomyces aureofaciens

Exemplary protocol of a biotransformation for the production of the compound (Ia) from the compound (IJa).

The method of Example 2 was used to prepare the production cultures using 50 μl spore suspension of the strain Streptomyces aureofaciens instead of the strain S. djakartensis NRRL B-12103.

Example 8

Biotransformation with Streptomyces djakartensis

Exemplary protocol of a biotransformation for the production of Spinosyn A with a 1-hydroxyethyl residue in the C-21 position [compound of the general

Formula (I) in which R ^{1 stands} for an amino sugar of the formula Ia, AB stands for the group -HC = CH- and D stands for the Grappe -CO-R ² , wherein R ^{2 stands} for a sugar of the formula 2a] ,

20 ml of R5A medium (R5A-

Medium: see Example 2) in 100 ml Erlenmeyer flasks with 50 μl spore suspension of S. djakartensis NRRL B-12103 inoculated. Before inoculation, the medium was sterilized at 121 ° C and 1.1 bar pressure for 20 minutes. The cultures were incubated at 28 ° C and 200 rpm. After 48 hours of incubation, 2 mg of Spinosyn A (100 μl of a stock solution of 10 mg / ml in methanol) were added. The biotransformation was stopped after 96 hours. The cultures were centrifuged off (10 minutes, 4000 rpm) and the supernatant was mixed with the same volume of methanol. Example 9

Biotransformation with Streptomyces dj kartensis

Exemplary protocol of a biotransformation for the production of 17-pseudospinosyn aglycon A with a 1-hydroxyethyl residue in the C-21 position [compound of the general formula (I), in which R ^{1 is} hydrogen, AB is the Group -HC-CH- or -HC = C (CH ₃ ) - and D stands for Grappe -CO-R ² , wherein R ^{2 stands} for a sugar of the formula 2a].

To prepare the production cultures, 20 ml of R5 A medium (R5 A medium: see Example 2) were inoculated in 100 ml Erlenmeyer flasks, each with 50 μl spore suspension of S. djakartensis NRRL B-12103. Before inoculation, the medium was sterilized at 121 ° C and 1.1 bar pressure for 20 minutes. The cultures were incubated at 28 ° C and 200 rpm. After 48 hours of incubation, 2 mg of 17-pseudospinosyn aglycone A or D (100 μl of a stock solution of 10 mg / ml in methanol) were added. The biotransformation was stopped after 96 hours. The cultures were centrifuged off (10 minutes, 4000 rpm) and the supernatant was mixed with the same volume of methanol.

Example 10

Isolation of the compound (la) from the biotransformation with S. djakartensis NRRL B-12103

Exemplary protocol for processing the culture supernatants and enrichment rank of the compound (la).

35 ml of the culture supernatant mixed with methanol from Example 2 were concentrated to about 20 ml and 10 ml of water were added. It was extracted twice with 10 ml of ethyl acetate each time, the combined organic phases were evaporated to dryness and the residue was taken up in 400 μl of methanol. An aliquot of this

Solution was analyzed by HPLC / MS (Example 11). Example 11

Analytical HPLC / / MS

Exemplary protocol for the analysis of the processed culture supernatants using

HPLC / UV / MS

An aliquot of the processed culture supernatant of the biotransformation with S. djakartensis (Example 2) was on a reversed-phase HPLC column (2.1 x 250 mm) with a gradient of water, mixed with ammonium acetate

(25 mmol / l), and methanol, mixed with ammonium acetate (25 mmol / l), chromatographed at a flow rate of 250 μl / minute. Detection is carried out with UV (245 nm) and electrospray (positive) mass spectrometry on a quadrupole mass spectrometer.

Compound (la) has a molecular weight of 418 daltons and is detected under these conditions as [M + NH ₄ ] ⁺ ion at m / z = 436. The retention time is approx. 32.5 minutes shorter than that of the compound (Ua) at approx. 37.5 minutes.

Example 12

Extraction and preparative presentation of the compound (la) from shake cultures of the biotransformation with S. djakartensis NRRL B-12103

Fifteen 20 ml cultures of the strain (S. djakartensis) in 100 ml Erlenmeyer flasks were grown according to the procedure described in Example 2 and the

Culture supernatants combined to work up compound (la). The combined culture supernatants were worked up analogously to Example 11. The residue was taken up in about 3 ml of methanol. The compound (Ia) was isolated by chromatography on an analytical reversed-phase HPLC column (4.6 × 250 mm) with a gradient of 25 mmol / 1 ammonium acetate in water and

25mmol / l ammonium acetate in methanol. An aliquot of 100 μl per run injected. UV detection was carried out at 245 nm. The fractions were collected manually, combined and evaporated to dryness. The yield was about 1 mg.

Example 13

Structure elucidation of the connection (la)

The preparatively isolated compound (Ia) was recorded in CD ₃ OD and examined by nuclear magnetic resonance spectroscopy (NMR). 1H-NMR, COZY, TOCSY, HSQC and HMBC spectra were recorded. The results are summarized in the following table.

NMR data of compound (Ia) in CD ₃ OD (500 MHz)

Position δc 5H Mult. / [Hz] Int. [ppm] [ppm]

1 174 - - -

2 35 2.48 dd 14/3 1H

3.12 dd 14/5 1H

3 49 2.93 ddd 10/5/2 1H

4 43 3.49 m 1H

5 129 5.84 ddd 10/3/3 1H

6 131 5.91 d br. 10 1H

7 42 2.23 m 1H

8 41 1.46 m 1H

1.83 dd 13/7 1H

9 73 4.34 m 1H

10 40 1.24 m 1H

2.34 m 1H

11 47 0.94 m 1H

12 51 2.85 m 1H

13 150 7.00 s br. 1H Position δc δ _H mult. / [Hz] Int. [ppm] [ppm]

14 146 - -

15 206 - -

16 50 3.22 m 1H

17 73 3.49 m 1H

18 36 1.43 m 1H

1.48 m 1H

19 23 1.28 m 1H

1.71 m 1H

20 27 1.48 m 1H

1.63 m 1H

21 79 4.70 ddd 10/5/2 1H

22 70 3.65 m 1H

23 18 1.04 d 7 3H

24 16 1.15 d 7 3H

Examples 14

Analytical detection of the hydroxylated spinosyn A

20 ml of the culture supernatant of the biotransformation from Example 8 mixed with methanol were adjusted to pH 5 with 0.01 N NaOH solution and to about 5 ml concentrated aqueous residue. This was extracted twice with 5 ml of ethyl acetate each. The combined organic phases were evaporated to dryness in a stream of N ₂ and taken up in 200 μl of methanol. An aliquot of this extract was examined using LC / MS and LC / MS / MS on a tandem mass spectrometer with electrospray positive ionization.

In the LC / MS chromatogram of the extract, a peak appears at 40.0 minutes with [M + H] ⁺ m / z = 748.5 in low concentration. The daughter ion spectrum of this ion has a fragment with m / z = 142, which is characteristic of the cleavage of a forosamine unit.

Examples 15

Analytical detection of the hydroxylated 17-pseudo-spinosyn-

aglycone

20 ml of the culture supernatant of the biotransformation from Example 9 mixed with methanol were adjusted to pH 5 with 0.01 N NaOH solution and concentrated to about 5 ml of aqueous residue. This was extracted twice with 5 ml of ethyl acetate each. The combined organic phases were evaporated to dryness in a stream of N ₂ and taken up in 200 μl of methanol. An aliquot of this

Extracts were examined using LC / MS and LC / MS / MS on a tandem mass spectrometer with electrospray positive ionization.

At 38.8 minutes, a peak with [M + NH ₄ ] ⁺ m / z = 624.4 appears in low concentration in the LC / MS chromatogram of the extract. This corresponds to a hydroxylated product from 17-pseudo-spinosyn A aglycone. The daughter ion spectrum of this ion has a fragment with m / z = 189, which is characteristic of the cleavage of a trimethylrhamnose unit. Example 16

Characterization of the strains used

a) Streptomyces djakartensis NRRL B-12103: This strain was certified as a niddamycin producer by the Agricultural

Research Service Culture Collection (1815 N. University Street, Illinois 61604, U.S.A.) under accession number NRRL B-12103. The strain description can be found in US 3646194. The culture was again on 06.06.2001 under the deposit number DSM 14327 at the German Collection of Microorganisms and Cell Cultures GmbH

(DSMZ), Mascheroder Weg lb, D-38124 Braunschweig, Germany, in accordance with the provisions of the Budapest Treaty.

b) Streptomyces griseofuscus DSM 40191:

This strain was obtained as Bundlin A, B, Moldicidin A and pentamycin producer from the German Collection of Microorganisms and Cell Cultures (Mascheroder Weg lb, D-38124 Braunschweig, Germany) under accession number 40191. The culture was again on 06.06.2001 under the deposit number DSM 14330 with the German

Collection of microorganisms and cell cultures GmbH (DSMZ), Mascheroder Weg lb, D-38124 Braunschweig, Germany, in accordance with the provisions of the Budapest Treaty.

c) Streptomyces caelestis DSM 40084:

This strain was obtained as a caelesticetin producer from the German Collection of Microorganisms and Cell Cultures (Mascheroder Weg lb, D-38124 Braunschweig, Germany) under accession number 40084. The culture was again on 06.06.2001 under the deposit number 14328 at the German Collection of Microorganisms and Cell Cultures GmbH (DSMZ), Mascheroder Weg lb, D-38124 Braunschweig, Germany, in accordance with the provisions of the Budapest Treaty.

d) Streptomyces antibioticus ATCC 11891: This strain was a Caelesticetin producer from the American Type

Culture Collection (10801 University Boulevard, Manassas, VA 20110-2209, USA) under the accession number ATCC 11891. The culture was again on 06.06.2001 under the deposit number DSM 14329 at the German Collection of Microorganisms and Cell Cultures GmbH (DSMZ), Mascheroder Weg lb, D-38124 Braunschweig, Germany, in

In accordance with the provisions of the Budapest Treaty.

e) Streptomyces griseus DSM 40937: This strain was obtained from the German Collection of Microorganisms and Cell Cultures (Mascheroder Weg lb, D-38124 Braunschweig, Germany) under accession number 40937. The culture was again on 06.06.2001 under the deposit number DSM 14331 at the German Collection of Microorganisms and Cell Cultures GmbH (DSMZ), Mascheroder Weg lb, D-38124 Braunschweig, Germany, in

In accordance with the provisions of the Budapest Treaty.

f) Streptomyces aureofaciens DSM 46447: This strain was obtained as a tetracycline producer from the German Collection of Microorganisms and Cell Cultures (Mascheroder Weg lb, D-38124 Braunschweig, Germany) under accession number 40084. The culture was again on 06.06.2001 under the deposit number DSM 14332 at the German Collection of Microorganisms and Cell Cultures GmbH (DSMZ), Mascheroder Weg lb, D-38124 Braunschweig, Germany, in accordance with the provisions of the Budapest Treaty.

Example 17 Preparation of the starting compounds

Spinosyn A-Aglykon (Ila)

The spinosyn A-aglycon (Ila) [compound of the general formula (II), in which Rl is hydrogen, AB is the group -HC = CH- and D is the group C-OH] was described in WO 01/16303 Tracer ^® manufactured.

9-Keto-spinosyn A-Aglykon (Ilb)

The 9-keto-spinosyn A-aglycone (Ilb) [Compound of the general formula (IT), in which R is hydrogen, AB is the group -HC = CH- and D is the group C = O] was by pyridinium dichromate oxidation made from compound (Ila):

46.55 g (115.6 mmol) of the Spinosyn A aglycon (Ila) were added under inert gas

1100ml abs. Dissolved dichloromethane and mixed with 43.51 g (115.6 mmol) of pyridinium dichromate. After 4 hours of stirring at 25 ° C. and addition of 900 ml of diethyl ether, the chromium salts which had precipitated were filtered off and the filtrate was concentrated in vacuo. Column chromatography on silica gel (eluent: cyclohexane / ethyl acetate 1: 1, then 100% ethyl acetate) gave 3.68 g 9.17-

Diketospinosyn aglycone and 23.40 g of an approx. 9: 1 mixture of spinosyn A aglycone (Ila) and 17-keto-spinosyn aglycone (above) in addition to 11.74 g of recovered spinosyn A aglycone (Ila). By recrystallization of the mixture in cyclohexane / ethyl acetate, the 9-keto-spinosyn A-aglycone (above) is enriched to> 98%. 20.78 g of 9-keto-spinosyn A-aglycon (Ilb) are obtained as colorless crystals. TLC: R _f (SiO ₂ , ethyl acetate) = 0.44-1 H NMR: CDC1 ₃ , δ = 6.77 (s, 13-H); 5.97 (d, 6-H); 5.88 (m, 5-H); 4.72 (m, 21-H); 3.69 (m, 17-H) et al. - LC / ESI-MS: m / z = 401 (25%) [M] ⁺ , 289 (100%).

Diketospinosyn aglycone: TLC: R {(SiO ₂ , ethyl acetate) = 0.64. - 1H NMR: CDC1 ₃ , δ = 6.92 (s, 13-H); 5.97 (d, 6-H); 5.87 (m, 5-H); 4.85 (m, 21 -H); 4.25 (q, 16-H) inter alia - LC / ESI-MS: m / z = 399 (100%) [M + H] ⁺ .

Spinosyn A

To prepare Spinosyn A, the procedure described in WO 01/16303 was first used. The resulting Spinosyn A / D mixture was separated by chromatography on a preparative reversed-phase column (250 x 8 mm). Water with 25 mmol / L ammonium acetate (A) and methanol with 25 mmol / L ammonium acetate (B) were used as eluents. The elution was carried out with a

Gradients from 60% B to 100% B in 35 minutes. The flow rate was 3 ml / minute. The separated substances were detected with a UV detector at 242 nm and automatically fractionated. Spinosyn A eluted at approximately 31 minutes and Spinosyn D at approximately 33 minutes. The combined fractions of Spinosyn A from several injections were evaporated on a rotary evaporator in vacuo to the aqueous residue. This aqueous solution was subjected to freeze drying and Spinosyn A was obtained as a white solid.

17-pseudo-spinosyn A / D aglycon The 17-pseudo-spinosyn A / D aglycon was prepared as described in WO 01/16303. FOR THE PURPOSES VC43 PATENT PROCESS

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Claims

claims:

Nerher for the preparation of compounds of general formula (I),

in which

D for the group

or stands

R ^{1 represents} hydrogen or an amino sugar, and

R> 2 represents hydrogen or a sugar,

characterized in that compounds of the general formula

(Π),

in which

AB, D and R ^{1 have} the meanings given above,

A method according to claim 1, characterized in that compounds of general formula (II) are used,

in which case (1) that

AB stands for one of the following groups: -HC = CH-, -HC = C (CH3) -, -H ₂ C-CH ₂ - or -H ₂ C-CH (CH ₃ ) -, and

D for the group

stands,

R represents an amino sugar of the formula Ia,

1a and

R> 2 represents a sugar of the formula 2a,

me

2a

or in which case (2) that

AB stands for the group -HC = CH- or -H ₂ C-CH ₂ -, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the above formula la, and

R ^{2 represents} hydrogen or a sugar of the formula 2b, 2c, 2d, 2e or 2f,

me

2 B

me

2c

me

2d

me

2e

me

2f or in which case (3) that

AB stands for one of the following groups: -HC = CH-, -HC = C (CH3) - or -H ₂ C-CH -, and

D has the meaning given above, R represents hydrogen or an amino sugar of the formula Ib,

1b

and

R represents hydrogen or a sugar of the abovementioned formula 2a,

or in which case (4) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the above formula la, and

R represents a sugar of the formula 2g, 2h, 2i, 2j or 2k,

me

2g

me

2h

me

2i

me

2k

or in which case (5) that

AB stands for the group -HC = CH- or -H ₂ C-CH ₂ -, and

D has the meaning given above,

R represents an amino sugar of the above formula 1 a, and

R ^{2 represents} a sugar of the formula 21 or 2m,

me

21

me

2m

or in which case (6) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ , and

D has the meaning given above,

1c

and

R ^{2 represents} a sugar of the above-mentioned formula 2b, 2c, 2g or 2h, or a sugar of the formula 2n,

me

2n

or in which case (7) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the above formula la, and

R> 2 represents a sugar of the formula 20,

me

2o

or in which case (8) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} an aminosugar of the abovementioned formula Ib, and R ^{2 represents} a sugar of the abovementioned formula 2d, 2i or 2j, or a sugar of the formula 2p,

2p

or in which case (9) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} hydrogen or an amino sugar of the above formula Ic, and

R ^{2 represents} a sugar of the above formula 2i or 2p,

or in which case (10) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the formula Id, Le or Lf,

1d

1e

1f

and

R> 2 represents a sugar of the above formula 2a,

or in which case (11) that

A-B stands for the group -HC = CH-, and

D for the group stands, R ^{1 represents} hydrogen or an amino sugar of the above formula la.

3. The method according to claim 1 or 2, characterized in that compounds of the general formula (11) are used,

in which case (12) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and

D for the group

R ^{1 represents} an amino sugar of the formula 1 a, and

R ^{2 represents} a sugar of the formula 2a, 2g or 2h,

or in which case (13) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the formula 1a, and

R ^{2 represents} hydrogen or a sugar of the formula 2d, 2e, 21, 2m or 2o,

or in which case (14) that AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and

D has the meaning given above,

R ^{1 represents} hydrogen or an amino sugar of the formula Ib, and

R ^{2 represents} hydrogen or a sugar of the formula 2a,

or in which AB, D and R ^{1 are defined} as in case (11).

4. The method according to any one of claims 1 to 3, characterized in that compounds of the general formula (II) are used,

in which case (15) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ , and

D for the group

R ^{1 represents} an amino sugar of the formula 1 a, and

R ^{2 represents} a sugar of the formula 2a,

or in which case (16) that

A-B stands for the group -HC = CH-, and

R ^{2 represents} hydrogen or a sugar of the formula 2d, 21 or 2m,

or in which AB, D and R ^{1 are defined} as in case (11).

5. The method according to any one of claims 1 to 4, characterized in that compounds of the general formula (II) are used,

in which case (17) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and

D for the group

R ^{1 represents} an amino sugar of the formula Ia, and

R represents a sugar of the formula 2a,

or in which case (18) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} hydrogen, and

R ^{2 represents} hydrogen. Method according to one of claims 1 to 5, characterized in that a microorganism from the group of the Actinomycetes is used as the microorganism.

A method according to claim 6, characterized in that a microorganism from the genus Streptomyces is used as the microorganism from the group of Actinomycetes.

A method according to claim 7, characterized in that a microorganism from the genus Streptomyces is a strain from the species Streptomyces djakartensis, Streptomyces griseofuscus, Streptomyces caelestis, Streptomyces antibioticus, Streptomyces griseus or Streptomyces aureofaciens.

9. The method according to claim 8, characterized in that the strain has the characteristic features of the strains Streptomyces djakartensis DSM

14327, Streptomyces griseofuscus DSM 14330, Streptomyces caelestis DSM

14328, Streptomyces antibioticus DSM 14329, Streptomyces griseus DSM 14331 or Streptomyces aureofaciens DSM 14332.

10. The method according to any one of claims 1 to 9, characterized in that the compounds of the general formula (I) are isolated.

11. Compounds of the general formula (I),

in which

D for the group

or stands

R ^{1 represents} an amino sugar, and

R stands for a sugar.

12. Compounds according to claim 11, characterized in that

in the case (1) that

AB represents one of the following groups: -HC = CH-, -HC = C (CH ₃ ) -, -H ₂ C-CH ₂ - or -H C-CH (CH ₃ , and

D for the group

R ^{1 represents} an amino sugar of the formula 1a,

1a and

R represents a sugar of the formula 2a,

me

2a

or in the case (2) that

AB stands for the group -HC = CH- or -H ₂ C-CH ₂ -, and

D has the meaning given above,

R ^{1 stands} for an amino sugar of the above formula 1 a, and

R represents hydrogen or a sugar of the formula 2b, 2c, 2d, 2e or 2f,

2 B

me

2c

me

2d

me

2e

me

2f

or in the case (3) that

AB stands for one of the following groups: -HC = CH-, -HC = C (CH ₃ ) - or -H ₂ C-CH ₂ -, and

D has the meaning given above,

R ^{1 represents} hydrogen or an amino sugar of the formula Ib,

1 bunch

R represents hydrogen or a sugar of the abovementioned formula 2a,

or in the case (4) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the above formula la, and

R represents a sugar of the formula 2g, 2h, 2i, 2j or 2k,

me

2g

me

2h

me

21

2y

me

2k

or in the case (5) that

AB stands for the group -HC = CH- or -H ₂ C-CH ₂ -, and

D has the meaning given above,

R represents an amino sugar of the above formula 1 a, and

R represents a sugar of the formula 21 or 2m,

me

21

me

2m

or in the case (6) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and

D has the meaning given above,

1c

and

R represents a sugar of the abovementioned formula 2b, 2c, 2g or 2h, or a sugar of the formula 2n,

me

2n

or in the case (7) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the above formula la, and

R> 2 represents a sugar of the formula 20,

me

2o

or in the case (8) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} an aminosugar of the abovementioned formula Ib, and

R for a sugar of the above formula 2d, 2i or 2j, or stands for a sugar of formula 2p,

or in the case (9) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} hydrogen or an amino sugar of the above formula Ic, and

R ^{2 represents} a sugar of the above formula 2i or 2p,

or in the case (10) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the above formula Id, le or an amino sugar of the formula If,

1d

1e

1f

and

R ^{2 represents} a sugar of the abovementioned formula 2a,

or in the case (11) that

A-B stands for the group -HC = CH-, and

13. Compounds according to claim 12, characterized in that

in the case (12) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and

D for the group

R ^{1 represents} an amino sugar of the formula 1 a, and

R ^{2 represents} a sugar of the formula 2a, 2g or 2h,

or in the case (13) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the formula 1a, and

R ^{2 represents} hydrogen or a sugar of the formula 2d, 2e, 21, 2m or

2o stands

or in the case (14) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and D has the meaning given above,

R ^{1 represents} hydrogen or an amino sugar of the formula Ib, and

R ^{2 represents} hydrogen or a sugar of the formula 2a,

or AB, D and R ^{1 are} as defined in case (11).

14. Compounds according to claim 12, characterized in that

in the case (15) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ , and

D for the group

R ^{1 represents} an amino sugar of the formula 1 a, and

R represents a sugar of the formula 2a,

or in the case (16) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} an amino sugar of the formula 1a, and R ^{2 represents} hydrogen or a sugar of the formula 2d, 21 or 2m,

or AB, D and R ^{1 are} as defined in case (11).

15. Compounds according to claim 12, characterized in that

in the case (17) that

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -, and

D for the group

R ^{1 represents} an amino sugar of the formula Ia, and

R ^{2 represents} a sugar of the formula 2a,

or in the case (18) that

A-B stands for the group -HC = CH-, and

D has the meaning given above,

R ^{1 represents} hydrogen, and

R ^{2 represents} hydrogen.

16. Compounds according to claim 12, characterized in that

AB stands for the group -HC = CH-, D for the group

stands,

R ^{1 represents} an amino sugar of the formula Ia, and

R represents a sugar of the formula 2a,

or in which

AB stands for the group -HC = CH- or -HC = C (CH ₃ ) -,

D has the meaning given above,

R ^{1 represents} hydrogen, and

R ^{2 represents} a sugar of the formula 2a,

or in which

A-B stands for a group -HC = CH-,

D has the meaning given above,

R ^{1 represents} hydrogen, and

R ^{2 represents} hydrogen.

17. Use of compounds according to any one of claims 11 to 16 for the preparation of new spinosyn derivatives.